WO2001070380A1

WO2001070380A1 - Lipid membrane, method for measuring membrane permeability, and method for screening

Info

Publication number: WO2001070380A1
Application number: PCT/JP2001/002346
Authority: WO
Inventors: Kiyohiko Sugano
Original assignee: Chugai Seiyaku Kabushiki Kaisha
Priority date: 2000-03-23
Filing date: 2001-03-23
Publication date: 2001-09-27
Also published as: DE60104190D1; US20030111406A1; JP3954847B2; DE60104190T2; EP1266684B1; US6861260B2; EP1266684A1; AU2001239564A1; ATE270581T1; EP1266684A4

Abstract

A membrane comprising a lipid and an unsaturated hydrocarbon having seven to nine carbon atoms. The membrane exhibits a permeability for a substance which is high and has a excellent correlation with the permeability of a drug, and thus suitable for use in rapid determination of permeability.

Description

TECHNICAL FIELD Lipid membrane, method for measuring membrane permeability and screening method

The present invention relates to a lipid membrane, and more particularly, the present invention relates to a lipid membrane that can be used for measuring the permeability of a substance. Background art

2. Description of the Related Art Conventionally, lipid membranes have been used as sensors or detection membranes of measurement devices, and are used for detecting or measuring various substances such as metal ions, cyanide ions, alcohols, and enzymes.

In the living body, transfer and metabolism of substances and energy, information transmission, and the like are performed through a biological membrane. A lipid membrane is also used to evaluate the permeability of the biological membrane.

For example, lipid membrane permeability of a drug is closely related to gastrointestinal absorption in an oral preparation and tissue transportability in a living body, and is an important property in drug development. In addition, evaluation of membrane permeability is important not only for drugs but also for substances that are harmful to living organisms (eg, toxic substances, carcinogenic substances, etc.).

When measuring the permeability of a substance in a living body in vitro, it is necessary to use a membrane having a substance permeability that is highly correlated with the permeability of a biological membrane in vivo. In addition, processing speed and cost are important in the industrial application of such membranes. For example, when used for drug permeability evaluation in the development of pharmaceuticals, a high processing speed is required because the number of compounds handled is very large.

Conventionally, in vitro measurement of a substance's membrane permeability includes a method using an isolated organ and a method using cells derived from gastrointestinal epithelium. However, these methods have a problem in measuring many substances quickly due to their slow processing speed. _{C In} addition, a method of measuring membrane permeability by creating an artificial lipid membrane in a 96-well plate (KA NS Y, Man fred, SENNER, Franck, GUBERNATO R, Kl au s; Jou rna lof Med ici na l Chemis ry 1998, 41, 1007-1010), etc., and such a lipid membrane is composed of a lipid and an organic solvent. This measurement method has features such as the ability to measure many substances at the same time, and the running cost is low because only a small amount of substances are used. However, the membrane used in the disclosed technique has not only a poor correlation with the permeability of the substance of the biological membrane in vivo, but also the low permeability of the substance, which makes it difficult to evaluate a low-permeability substance. However, there were drawbacks such as a long measurement time.

As a membrane for measuring the membrane permeability of a substance, a method in which a membrane component collected from a living body (rat) is dissolved in n-decane (I NUI, Ken-ichi, TABARA, Katsue) , HIRI, Ryohei, KANEDA, Akemi, MURAN ISHI, Shozo, SEZAKI, Hitoshi; Jou rna lof Pharrmacy and Pharmacology, 1977, 29, 22-26), etc. It has been known. This membrane has a characteristic that it has a good correlation with the permeability of substances in biological membranes, but has the drawbacks that it is difficult to evaluate low-permeability substances due to the low permeability of substances and that the measurement time is long. .

In addition, a membrane using dodecane, phosphatidylcholine, and 1,9-decadiene (KANSY, Manfred, SENNER, Frank, GUBERNATOR, Klaus; JorunalofMedicini nal Chemistry 1998, 41) , 1007-1010) are known, but the measurement time is long due to the low permeability of the substance, and it is difficult to distinguish between the low permeability substances. There were drawbacks such as poor correlation. Disclosure of the invention

As mentioned above, the membranes that have been used to measure the permeability of substances to date have long measurement times due to low permeability of substances, difficult to distinguish between low permeability substances, There are drawbacks such as poor correlation with the permeability of the substance. An object of the present invention is to provide a lipid membrane which overcomes these drawbacks, has a high substance permeability, has a high correlation with the substance permeability of a biological membrane, and is suitable for a rapid measurement method. The present inventors have conducted intensive studies and found that by using an unsaturated hydrocarbon having 7 to 9 carbon atoms, it is possible to obtain a lipid membrane having a high substance permeability and suitable for a rapid measurement method. This led to the completion of the present invention.

Furthermore, the use of unsaturated hydrocarbons having 7 to 9 carbon atoms, substances having a negative charge near neutrality, and lipids having a negative charge near Z or neutrality makes the drug highly transparent and in vivo. The present inventors have found that a lipid membrane suitable for a rapid measurement method can be obtained, which has a high correlation with the drug permeability of a biological membrane in the present invention, and has completed the present invention.

That is, the present invention provides a lipid membrane comprising an unsaturated hydrocarbon having 7 to 9 carbon atoms and a lipid.

It is preferable that the lipid membrane further contains a substance having a negative charge near neutrality. Alternatively, it is preferable that the lipid has a negative charge near neutrality.

Preferably, the unsaturated hydrocarbon having 7 to 9 carbon atoms is hebutadiene, octadiene or nonadiene. More preferably, it is 1,6-butadiene, 1,7-butadiene or 1,8-nonadiene.

Other elements may be added to the lipid membrane of the present invention. For example, a carrier for transporting a specific substance (specific transporter) may be added.

Further, the present invention provides a measuring method, comprising a step of measuring the membrane permeability of a substance using the above-mentioned lipid membrane.

Further, the present invention provides a membrane permeability measurement kit comprising an unsaturated hydrocarbon having 7 to 9 carbon atoms and a lipid. Such a measurement kit may further include a support (for example, a filter paper) that can support a lipid membrane composed of an unsaturated hydrocarbon having 7 to 9 carbon atoms and a lipid.

The present invention provides a method of screening a substance, comprising: a step of measuring the membrane permeability of a substance using the above-mentioned lipid membrane; and a step of selecting a substance having a predetermined membrane permeability. I do.

Further, the present invention provides a screening kit comprising an unsaturated hydrocarbon having 7 to 9 carbon atoms, a lipid, and an instruction for use.

Further, the measurement method of the present invention may be combined with a method of measuring active transport of a membrane, and a screening method including such a method and a kit used therefor are also provided. It is included in the scope of the present invention.

The lipid membrane of the present invention can be applied not only to the evaluation of the permeability of a substance through a biological membrane but also to a sensor or a measuring device for a chemical substance or the like.

BEST MODE FOR CARRYING OUT THE INVENTION

The disclosures of Japanese Patent Application No. 2000-82177 and Japanese Patent Application No. 2000-184973, which are the applications on which the priority claimed in the present application is based, are incorporated herein by reference.

The unsaturated hydrocarbon having 7 to 9 carbon atoms used in the present invention may be linear or branched, and includes, for example, hebutadiene, octadiene and nonadiene.

Examples of hebutadiene include (Z) —1,3-heptadiene, (Z) —1,4-heptadiene, (Z) —1,5-heptadiene, 1,6-heptadiene, and (E) — 1,3-butadiene, (E) — 1,4-butadiene, (E) — 1,5—butadiene, (2 Z, 4 Z) -2,4 butadiene, (2Z, 5 Z) 1,2-butadiene, (2Z, 4E) -2,4-butadiene, (2Z, 5E) -2,5 butadiene, (2E, 4Z) 1,2,4-butadiene, (2E, 5Z)-2,5-butadiene, (2E, 4E) -2,4 butadiene, (2E, 5E) -2,5_butadiene, (3Z, 5Z) 1,3-butadiene, (3Z, 5E)-3,5_butadiene, (3E, 5Z) 1,3,5-heptadiene, (3E, 5E) —3,5— Hebutadiene is preferred, and 1,6-hepnogene is preferred.

For example, (Z)-1, 3—Taktagen, (Z) — 1,4 — Taktagen, (Z) — 1,5 — Taktagen, (Z)-1,6 — Oktazyne, (E) — 1,3—year-old, (E) — 1,4-octadiene, (E) — 1,5-octadiene, (E) — 1,6-octadiene, 1,7-octadiene, (2 Z, 4 Z) -2,4-octadiene, (2 Z, 5 Z) one 2,5 _ octadiene, (2 Z, 6 Z) -2,6-o-octadiene, (2 Z, 4E) one twenty four —Octadjen, (2Z, 5E) -2,5—Octadjen, (2Z, 6E) -1 2,6—Octadjen, (2E, 4Z) -1 2,4-—Octadjen, (2E, 5Z) One 2,5-octactogen, (2E, 6 Z) -2,6-octactogen, (2E, 4 E) one 2,4-octactogen, (2E, 5 E) -2,5-octoctene, (2E , 6E)-2,6-octane, (3Z, 5Z) 1-3,5-octane, (3Z, 5E) 1,3,5-octane, (3E, 5Z) 1-3 , 5-octadiene, (3E, 5E) — 3,5-octadiene, and 1,7-octadiene is preferred.

As nonagen, for example, (Z) — 1, 3-Nonagen, (Z) —1,

(Z) one 1 (ヽ

4 1 P.-, "

Noah 丄, 乙) 丄, わ

(ヽヽ,, y ,,

L) 1 1, 7- Λ

\ ϊ ^) 丄, d— 丄, 4―

-N, (E)-1,5-Nonagen, (E)-1, 6-No ゝ (E, 丫

)-1, 7—nona, 1, (2 Z, 4 Z) one 2,4-nona

(2 Z, 5 Z) — 2,5-Nonagen, (2 Z, 6 Z) one 2,6-Nona _,

(2 Z, 7 Z)-2, (2 Z, 4E) one 2, 4-nona

(2 Z, 5 E) 1-2, (2 Z, 6 E) -2, 6-Nona

(2 Z, 7E)-2, (2E, 4Z) One 2, 4-nona

,-,

(2 E, 5 Z) 1-2, (2E, 6 Z) 1-2, 6 —Nona

(2 E, 7 Z) one 2, (2E, 4E) one 2, 4-nona

,

(2 E, 5E)-2, (2E, 6 E) -2, 6 —Nona

(2E, 7E)-2, (3 Z, 5 Z) -3, 5-nona

(3 Z, 6 Z)-3, 6-nonagen, (3 Z, 5 E) -3, 5-nona

,,,,, z (3 Z, 6E)-3, (3E, 5 Z) one 3,5-nona

(3 E, 6 Z)-3, (3E, 5 E) 1 3, 5-nona

,,hand

(3 E, 6 E) One is 3,6-nonagen, preferably 1,8-one.

One or a mixture of two or more of these butadiene, octadiene and nonadiene may be used as an unsaturated hydrocarbon having 7 to 9 carbon atoms. The lipid membrane of the present invention comprises, as an organic solvent, only an unsaturated hydrocarbon having 7 to 9 carbon atoms. It may contain a mixture of another organic solvent and an unsaturated hydrocarbon having 7 to 9 carbon atoms.

Examples of the lipid used in the present invention include saturated fatty acids, unsaturated fatty acids, phospholipids, and cholesterol.

Examples of the saturated fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, and behenic acid.

Examples of unsaturated fatty acids include palmitoleic acid, linolenic acid, linoleic acid, oleic acid, and arachidonic acid.

Examples of the phospholipid include phosphatidylcholine, phosphatidylglycerol, phosphatidylinosyl, phosphatidylserine, phosphatidylethanolamine, sphingomyelin and the like.

One of these lipids or a mixture of two or more thereof may be used.

Examples of the substance having a negative charge near neutrality used in the present invention include stearic acid, phosphatidylserine, phosphatidylinositol and the like. These may be used alone or in combination of two or more.

The ratio of lipid to unsaturated hydrocarbon having 7 to 9 carbon atoms (lipid weight / weight of unsaturated hydrocarbon having 7 to 9 carbon atoms) in the lipid membrane of the present invention is preferably 0.1 to 20%. It is more preferably 1 to 10%, particularly preferably 1 to 2%.

When the lipid membrane of the present invention contains a substance having a negative charge near neutrality, it should be contained at a ratio of about 0.2 mmo1 / L to about 50 mmo1 / L with respect to the entire lipid membrane. And more preferably in a ratio of about 2 mmo 1 ZL to about 25 mmo 1 / L. Here, “near neutral” generally means a range of pH 5.0 to 9.0, preferably pH 5.5 to 8.0, and more preferably pH 6.0 to 7.0. It is 5.

Further, "having a negative charge near neutral" means having a negative charge in a predetermined solvent near neutral.

The lipid membrane of the present invention may contain components other than those described above, and these components can be appropriately selected in consideration of a substance to be evaluated, characteristics of a living body or a tissue, and the like. For example, it may contain a carrier (specific transporter) for transporting a specific substance. The thickness of the lipid membrane of the present invention is appropriately selected depending on the substance to be evaluated, the characteristics of a living body or a tissue, and the like.

The lipid membrane of the present invention may be formed on any support. As such a support, a porous sheet-like material or a film-like material such as a filter paper is preferable. As a material, a hydrophobic substance is preferable, and for example, PTFE (polytetrafluoroethylene) and hydrophobic PVDF (polyvinylidene difluoride) can be used. Preferably, hydrophobic PVDF is used. Those having a pore size of 0.01 to 20 m, preferably 0.05 to: L0xm, more preferably 0.1 to 5 m, and particularly preferably about 0.1 to 1 m are used.

Alternatively, the lipid membrane of the present invention may be formed so as to cover an opening of a pore having a diameter of about 0.5 to 2 mm, preferably about 1 mm.

The lipid membrane of the present invention is obtained by mixing the above-mentioned unsaturated hydrocarbon having 7 to 9 carbon atoms and lipid by a usual method, for example, KANSY, Manfred, SENNER, Franck, GUBERNATOR, Klaus It can be produced according to the method described in JournalofMedicinal Chemistry 1998, 41, 1007-1010.

In addition, the measurement method of the present invention includes a step of measuring the membrane permeability of a substance using the lipid membrane of the present invention. The lipid membrane used for the measurement is formed and stored prior to the measurement. Alternatively, it may be formed immediately before the measurement. When the lipid membrane is formed, it is preferably formed on a support, more preferably on a hydrophobic support. Place a solution containing the test substance on one side (referred to as A side) of the obtained lipid membrane, and place a solvent containing no test substance on the other side (referred to as B side). At this time, the solvent on the B side is preferably the same as the solvent of the test substance solution on the A side. After a lapse of a predetermined time, the amount of the test substance that has passed through the lipid membrane from side A to side B is measured. In addition to directly quantifying the amount of test substance that has permeated through the lipid membrane, the amount of test substance that has permeated through the lipid membrane is calculated by measuring the amount of test substance remaining on the B side without permeating through the lipid membrane May be. The lipid membrane may be installed in any direction with respect to gravity. For example, the lipid membrane may be installed parallel or perpendicular to gravity. Also, the direction in which the substance permeates is not particularly limited. For example, even if the direction is the same as the gravity, Or vertical.

For measurement, one container may be divided into two compartments using a lipid membrane.However, a container for the test substance solution and a container for the solvent without the test substance may be separately prepared. . For example, a cylindrical upper container having a filter at the bottom and a lower container having an open top can be used in combination. Preferably, the lower end of the upper container and the upper end of the lower container have the same size. Fill the lower container with a solvent that does not contain the test substance, place the upper container on the lower container, form the lipid membrane of the present invention on the filter, put the test substance solution in the upper container, and after a predetermined time elapse, lower container The amount of test substance in the sample may be measured. Set the upper container and the lower container so that the filler in the upper container is in contact with the solvent in the lower container. It suffices if the filter can be fixed between the upper and lower containers so that the liquid does not leak from the upper and lower containers with the filter sandwiched between the upper and lower containers. Rubber packing or the like may be used. Also, the filter need not necessarily be fixed to the bottom of the upper container.

In order to quickly measure the membrane permeability of a plurality of test substances, it is also preferable to use a well plate having a plurality of wells. Use a well plate with multiple wells as the lower container, fill each well with a solvent that does not contain the test substance, cover it with a porous membrane such as Filler paper, and then match each well. An upper plate (used as an upper container) with through holes at the position may be placed, a lipid membrane may be formed on the porous membrane in each through hole, and the test substance solution may be charged. Alternatively, a filter may be provided at the bottom of each through hole of the upper plate. For example, a chemoplate such as chemotaxis ¾ ||

In each case, the upper container may contain the solvent without the test substance and the lower container may contain the test substance solution.

Methods for quantifying substances permeating the lipid membrane include, for example, absorbance measurement, HPLC method, TLC (thin layer chromatography) method, GC-MS (gas chromatography mass spectrum) method, LC-MS (liquid chromatography-mass spectrum) Method, fluorescence method, NMR method, IR method, CE (capillary electrophoresis) method and the like can be used, but P method, HPLC method, and LC-MS method are preferably used.

In the present invention, KAN SY, Manf The membrane permeability of a substance was measured according to the method described in J. rn lof Medicinl Chemistry 1998, 41, 1007-1010. Besides, for example, the method described in THOMPSON, Michael, LENNOX R. Bruce, MCCLELLAND, RA, Ana lytical Chemistry, 1982, 54, 76-81, XIA NG, Ti an-xi It can be measured according to the method described in ang, ANDERSON, Bradley D., Journ lof Pharmaceu itical Science, 1994, 83, 1511 and the like. To screen a substance using the lipid membrane of the present invention, the membrane permeability of the test substance is measured as described above, and a test substance having a predetermined membrane permeability is selected. At this time, some reference substance may be used to select a substance having higher or lower membrane permeability than that substance. Alternatively, a test substance having the same membrane permeability as the substance may be selected.

The screening kit of the present invention is useful for performing such a screening method quickly and easily, and includes an unsaturated hydrocarbon having 7 to 9 carbon atoms, a lipid, and an instruction manual. Membrane support (eg, filter paper, preferably hydrophobic filter paper, etc.), gel plate used for measuring membrane permeability, solvent, reference material for measuring membrane permeability, carrier for transporting specific substances ( (Specific transporter), and a solvent for dissolving the test substance. Further, the screening kit of the present invention may include a lipid membrane that has already been formed in place of the unsaturated hydrocarbon having 7 to 9 carbon atoms and lipid, for example, a lipid formed on a support. A membrane may be included.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. For example, the thickness of the prepared lipid membrane, the lipid concentration, the pH of the buffer solution, and the like can be appropriately changed according to the substance to be evaluated, the characteristics of the living body or tissue, and the like. Example

Formation of lipid membrane

Transfer each well (volume 360 / xL) of a 96-well plate (Fa1con flat bottom plate 3072) to 50 mm o 1 L phosphorus containing 5% dimethyl sulfoxide at pH 6.5 to 7.0. After filling with an acid sodium buffer (hereinafter referred to as "buffer"), a filter plate (Millipore Multiscreen filter plate hydrophobic PVDF MAIPN4510, thickness 0.45 μm) was covered on the well plate. At this time, it was confirmed that no bubbles were present between the buffer solution of the well plate and the filter plate. Each substance shown in Table 1 (indicating the composition of the film) was dissolved in 1,7-octadiene at the ratios shown in Table 1. The membranes of Examples 1 and 2 were formed on a part of the filter by adding 4 to 5 L of the obtained 1,7-year-old kutadiene solution to each filter of each well.

The membrane of Example 3 was formed on a part of the filter by adding 4 to 5 zL of phosphatidylcholine: 1,6-butadiene = 2: 98 at the filter of each well. The membrane of Example 4 was formed on a part of the filter by adding 4 to 5 L of a solution of phosphatidylcholine: 1,7-octane = 2: 98 to each filter. The membrane of Example 5 was formed on a part of the filter by adding 4 to 5 × L of a solution of phosphatidylcholine: 1,8-nonagen: 2: 98 to each filter.

Also, phosphatidylcholine: cholesterol: 1,9-decadiene = 2:

A 1:97 solution was prepared, and 4 to 5 L of the solution was added to each filter, thereby forming a membrane of Comparative Example 1 on a part of the filter. By adding solutions 4 to 5 of phosphatidylcholine: 1,9-decadiene = 2: 98 to the filters of each well, the membrane of Comparative Example 2 was formed on the filter portion. table 1

Measurement of membrane permeability

As sample solutions, 100 to 200 zL (amount of added sample solution: Vdn) of 0.5 mmol ZL buffer solution of each compound shown in Tables 2 and 3 was prepared. This sample solution was added to each of the wells on which a film was formed as described above, and the plate was covered and left for 2 to 15 hours (transmission time: t). After removing the filter plate, 200 solutions were collected from each well of the lower well plate and used as a test solution.

The absorbance (ODac) of the obtained test solution (200 L) was measured at each wavelength of 250 to 450 nm (interval of 10 to 20 nm). As the standard solution, each of the sample solutions used in Example 1 or 3 (an undiluted standard solution) or a solution obtained by diluting the sample solution 4.8-fold (V / V) with a buffer solution (dilution) Standard solution). That is, the absorbance was measured in the same manner as in the test solution using 200 L of either the undiluted standard solution or the diluted standard solution (ODref), and the transmission coefficient was calculated according to the following equation.

Transmission coefficient (P) = _ 2.303 X (V _dn XV _ac / (V _dn XV _ac )) Z (SX t) X 1 og (1-f 1 ux% / l 00)

F 1 ux% = OD _ac / OD _ref XAX 100

OD _AC : Measured absorbance of test solution

OD _{RE F:} absorbance measurements of standard solution

V _ac : Volume of each well of the lower plate (360 L)

V dn added sample solution volume (100-200 L) A: VdnXVac (for undiluted standard solution)

1 (for diluted standard solution)

S: membrane area (0.266 cm ² )

t: Transmission time (sec)

Tables 2 and 3 show the obtained transmission coefficients (P).

Further, for Examples 1, 2 and Comparative Example 1, the following approximate curves obtained from the gastrointestinal absorptivity (Fa) of each compound in humans (described in the references * 1 to * 4) were used. The correlation coefficient (R) was calculated.

Approximate curve: Fa () = (1-exp (-aXP)) X 100

exp: expone n t i a l

a: Coefficient

氺 1: Matthew D. Wessel et al., J. Chem. Inf. Comput. Sci., 38, 1998, 726-735

* 2: Mehran Yazdanian et al., Pharm. Res., Vol. 15, No. 9, 1998, 1490-1494

* 3: Gerald K. McEvoy, AHFS, 1998

* 4: Manfred Kansy et al., J. Med. Chem., Vol. 47, No. 7, 1998, 1007-1010.

Table 2 shows the obtained correlation coefficient (R) and the gastrointestinal absorption (Fa) of each compound in humans.

Table 2 Compound name Example 1 卖 Example 2 Comparative example of upper J-tube | 7 11

(P) (P) (P) Chemical absorption pH 7.0 pH 6.5 pH 6.5 (Fa,%) Acebutrol 4.45E- 06 3.67E-06 1.91E-07 90 * ¹ Acetaminophen 8.17E-06 1.79E-06 4.98B-08 80 * ^1- Acyclovir 7.97E-08 8.89E-08 3.87E-08 20 * ² -amidoride 2.69E-06 6.73E-07 6.10E-08 50 * ³ Atenolol 8.02E-07 8.64E-07 4.18E-07 50 * ¹ Ceftriaxone 1.37E-06 2.25E-07 1.66E-08 1 4 Cefuroxime 1.60E-07 4.43E-08 2.76E-08 5 * ¹ Cloth thiazide 2.24E -06 2.71E-07 4.98E-08 13 * Ί cytarabine 8.89E-07 3.87E-08 3.87E- 08 20 * 3 doxycycline 7.07E-06 2.35E-05 1.52E- 06 95 * 3 Enarapuriru 8.02E-07 1.37E-06 1.09E-06 65 * ³ Furosemide 3.09E-06 9.02E-07 1.34E-07 61 * ¹ Guanabenes 1.95E-05 1.04E-05 4.14E-06 75 * ¹ Hydrochloride thiazide 1.34E -06 1.55E-06 8.64E-07 67 * ¹ Hydrocortisone 8.84E-06 1.44E-05 3.18E-06 91 * ¹ Metoprolol 9.42E-06 6.88E-06 7.49E-07 95 * ¹ Nadolol 3.28 E-06 1.14E-06 5.26E-07 35 * ¹ Naltrexone 1.12E-05 4.50 E-06 9.61E-06 9 ₆ * 3 year old cyclin 3.31E-06 5.87E-06 8.31E-07 6 ₀ * 3 Pindolol 1.10E-05 7.94E-06 2.71E-08 90 * ¹ plaque Roll 1.01E-06 1.53E-06 4.36E-07 100 * ¹ Pravastatin 2.69E-06 6.12E-07 7.21E-08 34 * ¹ Proforce inamide 4.58E-06 3.89E-06 1.17E-07 85 * ³ Quinidine 2.56E-05 1.7E-05 4.51E-06 80 * ¹ Ranitidine 1.2E-06 2.19B-06 1.34E-07 50 * ¹ Sulfasalazine 3.24E-06 1.07E-06 3.12E-07 65 * ¹ Sulpiride 1.34E-06 2.23E-06 8.33E-08 35 * ⁴ Tetracycline 2.65E-06 7.62E-06 1.Similar -06 77.5 * ³ Timolol 9.13E-06 1.19E-05 1.05E-06 90 * ¹ human gastrointestinal absorption and P 0.798 0.847 0.534

Correlation coefficient (R) Table 3

Industrial applicability

ADVANTAGE OF THE INVENTION According to the present invention, a lipid membrane having high permeability of a substance, having a high correlation with the permeability of a biological membrane, and suitable for rapid measurement can be provided, and is extremely useful.

Claims

The scope of the claims

1. A lipid membrane comprising an unsaturated hydrocarbon having 7 carbon atoms and 9 and a lipid.

2. The lipid membrane according to claim 1, comprising a substance having a negative charge near neutrality.

3. The lipid according to claim 1, wherein the lipid has a negative charge near neutrality.

4. The lipid membrane according to any one of claims 1 to 3, wherein the unsaturated hydrocarbon having 7 to 9 carbon atoms is hebutadiene, octadiene or nonadiene.

5. The unsaturated hydrocarbon having 7 to 9 carbon atoms is 1,6-butadiene, 1, Ί-octadiene or 1,8-nonadiene. Lipid membrane.

6. A measurement method, comprising a step of measuring the membrane permeability of a substance using the lipid membrane according to any one of claims 1 to 5.

7. A membrane permeability measurement kit comprising an unsaturated hydrocarbon having 7 to 9 carbon atoms and a lipid.

8. A method comprising: measuring a membrane permeability of a substance using the lipid membrane according to any one of claims 1 to 5; and selecting a substance having a predetermined membrane permeability. Substance screening method.

9. A screening kit comprising an unsaturated hydrocarbon having 7 to 9 carbon atoms, a lipid, and instructions for use.